Splenocyte Preperation
Harvest mouse spleen and prepare a single cell suspension .
Use slides or a syrings to push the spleen through a cell strainer .
Pellet the cells by centrifugation ( 350 x g ) ; aspirate the supernatant .
Dilute the 10X Red Blood Cell Lysis Buffer to 1X working concentration with deionized water and resuspend the pellet in 5 ml of 1X Lysis Buffer .
Incubate on ice for 4 - 5 minutes with occasional shaking .
Stop the reaction by diluting the Lysis Buffer with 20 - 30 ml of 1X PBS .
Spin the cells ( 350 x g ) and discard the supernatant .
Resuspend the pellet in the appropriate buffer Count cells , adjust density , and proceed with cell staining procedures .
